CN101020895A - Method for preparing silage rich in conjugated linoleic acid by lactobacillus plantarum fermentation - Google Patents
Method for preparing silage rich in conjugated linoleic acid by lactobacillus plantarum fermentation Download PDFInfo
- Publication number
- CN101020895A CN101020895A CNA2007100196553A CN200710019655A CN101020895A CN 101020895 A CN101020895 A CN 101020895A CN A2007100196553 A CNA2007100196553 A CN A2007100196553A CN 200710019655 A CN200710019655 A CN 200710019655A CN 101020895 A CN101020895 A CN 101020895A
- Authority
- CN
- China
- Prior art keywords
- silage
- ancla01
- plant lactobacillus
- rich
- bacterial strain
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
- 239000004460 silage Substances 0.000 title claims abstract description 26
- 238000000034 method Methods 0.000 title claims abstract description 16
- 240000006024 Lactobacillus plantarum Species 0.000 title claims abstract description 10
- 235000013965 Lactobacillus plantarum Nutrition 0.000 title claims abstract description 10
- 229940072205 lactobacillus plantarum Drugs 0.000 title claims abstract description 10
- 238000000855 fermentation Methods 0.000 title claims description 12
- 230000004151 fermentation Effects 0.000 title claims description 12
- OYHQOLUKZRVURQ-IXWMQOLASA-N linoleic acid Natural products CCCCC\C=C/C\C=C\CCCCCCCC(O)=O OYHQOLUKZRVURQ-IXWMQOLASA-N 0.000 title abstract description 6
- JBYXPOFIGCOSSB-GOJKSUSPSA-N 9-cis,11-trans-octadecadienoic acid Chemical compound CCCCCC\C=C\C=C/CCCCCCCC(O)=O JBYXPOFIGCOSSB-GOJKSUSPSA-N 0.000 title abstract description 5
- 229940108924 conjugated linoleic acid Drugs 0.000 title abstract description 5
- 230000001580 bacterial effect Effects 0.000 claims abstract description 31
- 238000004321 preservation Methods 0.000 claims abstract description 5
- 241000186660 Lactobacillus Species 0.000 claims description 32
- 229940039696 lactobacillus Drugs 0.000 claims description 32
- 241000196324 Embryophyta Species 0.000 claims description 28
- 229960004232 linoleic acid Drugs 0.000 claims description 23
- OYHQOLUKZRVURQ-HZJYTTRNSA-N Linoleic acid Chemical compound CCCCC\C=C/C\C=C/CCCCCCCC(O)=O OYHQOLUKZRVURQ-HZJYTTRNSA-N 0.000 claims description 21
- JVTAAEKCZFNVCJ-UHFFFAOYSA-N lactic acid Chemical compound CC(O)C(O)=O JVTAAEKCZFNVCJ-UHFFFAOYSA-N 0.000 claims description 20
- 241000894006 Bacteria Species 0.000 claims description 17
- 239000007788 liquid Substances 0.000 claims description 15
- 235000014655 lactic acid Nutrition 0.000 claims description 10
- 238000002360 preparation method Methods 0.000 claims description 10
- 239000004310 lactic acid Substances 0.000 claims description 8
- 235000015097 nutrients Nutrition 0.000 claims description 8
- 239000003921 oil Substances 0.000 claims description 7
- 235000019198 oils Nutrition 0.000 claims description 7
- 235000020238 sunflower seed Nutrition 0.000 claims description 7
- 235000019483 Peanut oil Nutrition 0.000 claims description 6
- 235000019484 Rapeseed oil Nutrition 0.000 claims description 6
- 238000011081 inoculation Methods 0.000 claims description 6
- 239000000312 peanut oil Substances 0.000 claims description 6
- 102000004895 Lipoproteins Human genes 0.000 claims description 5
- 108090001030 Lipoproteins Proteins 0.000 claims description 5
- 235000015112 vegetable and seed oil Nutrition 0.000 claims description 5
- 239000008158 vegetable oil Substances 0.000 claims description 5
- 230000004913 activation Effects 0.000 claims description 4
- 238000007789 sealing Methods 0.000 claims description 2
- 230000002269 spontaneous effect Effects 0.000 claims description 2
- 230000007480 spreading Effects 0.000 claims description 2
- 238000003892 spreading Methods 0.000 claims description 2
- 240000008042 Zea mays Species 0.000 abstract description 7
- 235000005824 Zea mays ssp. parviglumis Nutrition 0.000 abstract description 7
- 235000002017 Zea mays subsp mays Nutrition 0.000 abstract description 7
- 235000005822 corn Nutrition 0.000 abstract description 7
- 235000019728 animal nutrition Nutrition 0.000 abstract description 4
- 238000009629 microbiological culture Methods 0.000 abstract 1
- 238000007792 addition Methods 0.000 description 10
- 239000002609 medium Substances 0.000 description 8
- 238000012360 testing method Methods 0.000 description 8
- VLKZOEOYAKHREP-UHFFFAOYSA-N n-Hexane Chemical compound CCCCCC VLKZOEOYAKHREP-UHFFFAOYSA-N 0.000 description 6
- 239000000243 solution Substances 0.000 description 6
- 238000002474 experimental method Methods 0.000 description 4
- 239000002054 inoculum Substances 0.000 description 4
- 230000000877 morphologic effect Effects 0.000 description 4
- 238000012216 screening Methods 0.000 description 4
- FRXSZNDVFUDTIR-UHFFFAOYSA-N 6-methoxy-1,2,3,4-tetrahydroquinoline Chemical compound N1CCCC2=CC(OC)=CC=C21 FRXSZNDVFUDTIR-UHFFFAOYSA-N 0.000 description 3
- 241000030538 Thecla Species 0.000 description 3
- 238000006243 chemical reaction Methods 0.000 description 3
- 238000011156 evaluation Methods 0.000 description 3
- 235000013305 food Nutrition 0.000 description 3
- 244000005700 microbiome Species 0.000 description 3
- 235000013336 milk Nutrition 0.000 description 3
- 239000008267 milk Substances 0.000 description 3
- 210000004080 milk Anatomy 0.000 description 3
- 230000035479 physiological effects, processes and functions Effects 0.000 description 3
- 239000002994 raw material Substances 0.000 description 3
- 238000009631 Broth culture Methods 0.000 description 2
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 description 2
- 238000003794 Gram staining Methods 0.000 description 2
- SIKJAQJRHWYJAI-UHFFFAOYSA-N Indole Chemical compound C1=CC=C2NC=CC2=C1 SIKJAQJRHWYJAI-UHFFFAOYSA-N 0.000 description 2
- 241001465754 Metazoa Species 0.000 description 2
- 238000012408 PCR amplification Methods 0.000 description 2
- 241000282849 Ruminantia Species 0.000 description 2
- 238000010521 absorption reaction Methods 0.000 description 2
- 239000002253 acid Substances 0.000 description 2
- 235000015278 beef Nutrition 0.000 description 2
- 239000008103 glucose Substances 0.000 description 2
- 239000004519 grease Substances 0.000 description 2
- 150000002632 lipids Chemical class 0.000 description 2
- 244000144972 livestock Species 0.000 description 2
- 239000000463 material Substances 0.000 description 2
- 239000007787 solid Substances 0.000 description 2
- 239000007921 spray Substances 0.000 description 2
- 239000000758 substrate Substances 0.000 description 2
- 229920001817 Agar Polymers 0.000 description 1
- 235000016068 Berberis vulgaris Nutrition 0.000 description 1
- 241000335053 Beta vulgaris Species 0.000 description 1
- 102000016938 Catalase Human genes 0.000 description 1
- 108010053835 Catalase Proteins 0.000 description 1
- 238000007400 DNA extraction Methods 0.000 description 1
- RWSOTUBLDIXVET-UHFFFAOYSA-N Dihydrogen sulfide Chemical compound S RWSOTUBLDIXVET-UHFFFAOYSA-N 0.000 description 1
- 108010010803 Gelatin Proteins 0.000 description 1
- 241000726221 Gemma Species 0.000 description 1
- 241000589516 Pseudomonas Species 0.000 description 1
- MUPFEKGTMRGPLJ-RMMQSMQOSA-N Raffinose Natural products O(C[C@H]1[C@@H](O)[C@H](O)[C@@H](O)[C@@H](O[C@@]2(CO)[C@H](O)[C@@H](O)[C@@H](CO)O2)O1)[C@@H]1[C@H](O)[C@@H](O)[C@@H](O)[C@@H](CO)O1 MUPFEKGTMRGPLJ-RMMQSMQOSA-N 0.000 description 1
- 241000219793 Trifolium Species 0.000 description 1
- MUPFEKGTMRGPLJ-UHFFFAOYSA-N UNPD196149 Natural products OC1C(O)C(CO)OC1(CO)OC1C(O)C(O)C(O)C(COC2C(C(O)C(O)C(CO)O2)O)O1 MUPFEKGTMRGPLJ-UHFFFAOYSA-N 0.000 description 1
- 238000009825 accumulation Methods 0.000 description 1
- 239000008272 agar Substances 0.000 description 1
- 238000000246 agarose gel electrophoresis Methods 0.000 description 1
- WQZGKKKJIJFFOK-PQMKYFCFSA-N alpha-D-mannose Chemical compound OC[C@H]1O[C@H](O)[C@@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-PQMKYFCFSA-N 0.000 description 1
- 230000001093 anti-cancer Effects 0.000 description 1
- 230000000259 anti-tumor effect Effects 0.000 description 1
- 230000008901 benefit Effects 0.000 description 1
- 230000033228 biological regulation Effects 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- 230000036983 biotransformation Effects 0.000 description 1
- 150000001720 carbohydrates Chemical class 0.000 description 1
- 235000014633 carbohydrates Nutrition 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- 239000003795 chemical substances by application Substances 0.000 description 1
- 235000013365 dairy product Nutrition 0.000 description 1
- 230000007812 deficiency Effects 0.000 description 1
- 235000005911 diet Nutrition 0.000 description 1
- 230000037213 diet Effects 0.000 description 1
- 235000014113 dietary fatty acids Nutrition 0.000 description 1
- 238000010790 dilution Methods 0.000 description 1
- 239000012895 dilution Substances 0.000 description 1
- 238000004090 dissolution Methods 0.000 description 1
- 239000012153 distilled water Substances 0.000 description 1
- 239000003814 drug Substances 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 230000002708 enhancing effect Effects 0.000 description 1
- 238000000605 extraction Methods 0.000 description 1
- 235000019197 fats Nutrition 0.000 description 1
- 229930195729 fatty acid Natural products 0.000 description 1
- 239000000194 fatty acid Substances 0.000 description 1
- 150000004665 fatty acids Chemical class 0.000 description 1
- 238000012262 fermentative production Methods 0.000 description 1
- 239000012634 fragment Substances 0.000 description 1
- 235000020604 functional milk Nutrition 0.000 description 1
- 239000007789 gas Substances 0.000 description 1
- 239000008273 gelatin Substances 0.000 description 1
- 229920000159 gelatin Polymers 0.000 description 1
- 235000019322 gelatine Nutrition 0.000 description 1
- 235000011852 gelatine desserts Nutrition 0.000 description 1
- 239000003292 glue Substances 0.000 description 1
- 239000001963 growth medium Substances 0.000 description 1
- 229910000037 hydrogen sulfide Inorganic materials 0.000 description 1
- 230000001900 immune effect Effects 0.000 description 1
- PZOUSPYUWWUPPK-UHFFFAOYSA-N indole Natural products CC1=CC=CC2=C1C=CN2 PZOUSPYUWWUPPK-UHFFFAOYSA-N 0.000 description 1
- RKJUIXBNRJVNHR-UHFFFAOYSA-N indolenine Natural products C1=CC=C2CC=NC2=C1 RKJUIXBNRJVNHR-UHFFFAOYSA-N 0.000 description 1
- 230000005764 inhibitory process Effects 0.000 description 1
- 230000031700 light absorption Effects 0.000 description 1
- 235000020778 linoleic acid Nutrition 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 238000005259 measurement Methods 0.000 description 1
- 235000013372 meat Nutrition 0.000 description 1
- 230000004060 metabolic process Effects 0.000 description 1
- 239000000178 monomer Substances 0.000 description 1
- 235000014571 nuts Nutrition 0.000 description 1
- 230000037361 pathway Effects 0.000 description 1
- 230000001681 protective effect Effects 0.000 description 1
- MUPFEKGTMRGPLJ-ZQSKZDJDSA-N raffinose Chemical compound O[C@H]1[C@H](O)[C@@H](CO)O[C@@]1(CO)O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO[C@@H]2[C@@H]([C@@H](O)[C@@H](O)[C@@H](CO)O2)O)O1 MUPFEKGTMRGPLJ-ZQSKZDJDSA-N 0.000 description 1
- 238000011084 recovery Methods 0.000 description 1
- 230000001105 regulatory effect Effects 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 238000000926 separation method Methods 0.000 description 1
- 238000012163 sequencing technique Methods 0.000 description 1
- 239000002904 solvent Substances 0.000 description 1
- 230000003068 static effect Effects 0.000 description 1
Images
Landscapes
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Fodder In General (AREA)
Abstract
一种植物乳杆菌发酵制备富含共轭亚油酸青贮饲料的方法属微生物和动物营养与饲料技术领域。本发明的菌株(Lactobacillus plantarumANCLA01)于2007年1月8日保藏于中国微生物菌种保藏管理委员会普通微生物中心，保藏登记入册的编号CGMCCNo.1906。植物乳杆菌ANCLA01系自玉米青贮中分离、筛选、纯化培育出来，命名为植物乳杆菌ANCLA01(Lactobacillus plantarum ANCLA01)。该植物乳杆菌ANCLA01应用于青贮饲料，可以提高青贮饲料中共轭亚油酸的含量，生产出富含共轭亚油酸的功能性青贮饲料。
The invention discloses a method for preparing silage rich in conjugated linoleic acid by fermenting Lactobacillus plantarum, which belongs to the technical field of microbiology and animal nutrition and feed. The bacterial strain of the present invention (Lactobacillus plantarum ANCLA01) was deposited in the General Microorganism Center of the China Microbiological Culture Collection Management Committee on January 8, 2007, and the preservation registration number was CGMCCNo.1906. Lactobacillus plantarum ANCLA01 is isolated, screened, purified and cultivated from corn silage, named Lactobacillus plantarum ANCLA01 (Lactobacillus plantarum ANCLA01). The plantarum ANCLA01 is applied to silage, can increase the content of conjugated linoleic acid in the silage, and produce functional silage rich in conjugated linoleic acid.
Description
Technical field
The invention belongs to microorganism and animal nutrition and feed technical field, be specifically related to the new bacterial strain of a kind of high yield conjugated linolic acid (CLA, Conjugated linoleic acid) and utilize its fermentative production to be rich in the functional silage of CLA.
Background technology
Anticancer, functions such as promotion is grown, inhibition accumulation of fat, enhancing body immunological competence that conjugated linolic acid (CLA) has can improve pig, the fowl speed of growth and lean ratio, are the research focuses of subjects such as current medicine, protective foods and Animal nutrition.U.S. NRC classifies CLA as unique animal source lipid acid with antitumous effect.There are 4 kinds of isomer at least in CLA, and " Cis-9, trans-11-linolic acid " is main among the CLA, the active a kind of isomer of tool.CLA in ruminating animal such as milk and beef, the mutton food more than 90% is " Cis-9, a trans-11-linolic acid ", is human topmost CLA natural origin.Therefore, the milk that CLA content is high is called as functional milk, and beef, mutton that CLA content is high are called as functional foodstuff.At present, people improve CLA content in milk from ruminants, the meat by two kinds of diet nutrient regulatory pathways: the one, and daily ration adds grease or changes daily ration adipose-derived, with the linoleic content of CLA metabolism substrate in the increase cud, and then improve CLA content in the livestock product; The 2nd, directly add CLA or ruminally-protected CLA (promptly crossing cud CLA).But because CLA costs an arm and a leg, it is too high to add cost in the feed, uses few in the production practice.Therefore, the animal nutrition and feed scholar is seeking other suitable, cost effective method that improves CLA content in the livestock product.
Summary of the invention
Technical problem to be solved by this invention is: the deficiency that is intended to overcome above-mentioned direct interpolation conjugated linolic acid and changes the daily ration lipid conformation, provide a kind of can the high yield conjugated linolic acid the new bacterial strain of plant lactobacillus and utilize its fermentative preparation to be rich in the method for conjugated linolic acid silage, to improve the content of conjugated linolic acid in the ruminant.
The new bacterial strain of plant lactobacillus of the present invention system is from separation from corn silage, screening, purifying and cultivate called after plant lactobacillus ANCLA01 (Lactobacillus plantarum ANCLA01).
The method that this plant lactobacillus fermentative preparation is rich in the conjugated linolic acid silage is: after earlier the strains A NCLA01 of this high yield conjugated linolic acid being spread cultivation in batches, again tieback makes the linolic acid fermentation in the Vegetable oil lipoprotein be converted into conjugated linolic acid in ensilings such as corn, Radix Dauci Sativae, clover and beet again.
Description of drawings
Fig. 1 is the photo that plant lactobacillus ANCLA01 of the present invention takes at microscopically;
Fig. 2 is a plant lactobacillus ANCLA01 bacterial strain pcr amplification band photo of the present invention;
Fig. 3 is the ability of different strains bio-transformation conjugated linolic acid;
Fig. 4 is the utilization of plant lactobacillus ANCLA01 to carbohydrate in the fermentation substrate.
Plant lactobacillus ANCLA01 of the present invention (Lactobacillus plantarum ANCLA01) has been carried out in accordance with regulations preservation: depositary institution: China Committee for Culture Collection of Microorganisms common micro-organisms center abbreviates CGMCC as; Address: China. Beijing. Zhong Guan-cun, Institute of Microorganism, Academia Sinica; Preservation date: on January 8th, 2007; The numbering CGMCCNo.1906 that preservation is registered on the books.
Embodiment:
1. materials and methods
1.1 material: corn silage is collected in Hefei City white Supreme Being's dairy industry Development Co., Ltd; Substratum: pH6.5-6.8, the MRS broth culture, the PY substratum, MRS solid medium, MRS semisolid medium, MRS liquid nutrient medium and benefit are added 0.1g (100ml)
-1The MRS liquid nutrient medium of LA.Substratum is with the preceding 20min that all sterilizes under 121 ℃ of high pressure steam.
1.2 main agents: LA and CLA: purity 〉=99%, available from Sigma company; Bacterial genomes DNA extraction test kit, PCR test kit and glue reclaim test kit, available from precious biotechnology (Dalian) company limited; Other reagent is analytical pure.
1.3 bacterial screening:
1.3.1 primary dcreening operation: gather choose immediately after the ensiling Oranoleptic indicator preferably the ensiling branches and leaves immerse in 37 ℃ of distilled waters, remove branches and leaves after ten minutes, draw fresh even soak solution 10mL and be added to 500ml and be equipped with in the Erlenmeyer flask of 300mL MRS broth culture.37 ℃ static increase bacterium 96h after, dilution spread plate method is toppled over the MRS agar plate, cultivates 48h in 37 ℃ of anaerobism behind four rides, picking list bacterium colony percutaneous puncture-inoculation is cultivated rear defences for 37 ℃ and is preserved in refrigerator in the MRS semisolid medium.
1.3.2 multiple sieve: primary dcreening operation is preserved bacterial classification after 37 ℃ of activation, and the inoculum size by 5% is inoculated in the band nut test tube (d18mmx160mm) that 10mL MRS liquid nutrient medium is housed, and wherein linoleic acid content is 0.1g (100mL) in the MRS liquid nutrient medium
-1N
2Drive away air, measure CLA behind the anaerobically fermenting 24h, produce the CLA bacterium with screening.
1.4 CLA measures: with the normal hexane is solvent, and the CLA standard specimen is made into the solution of different concns, is reference with the hexane, in 233nm place its absorption value of mensuration, is X-coordinate with CLA concentration (ug/mL), light absorption value is an ordinate zou, the drawing standard curve.(1998) method operations such as Jiang are pressed in the extraction of CLA in the fermented liquid.Afterwards, institute is obtained fatty acid extract 5mL n-hexane dissolution, with the fermentation broth extract of not inoculating is reference, with ultraviolet spectrophotometer 00-350nm scope interscan again, read the absorption value at charateristic avsorption band 233nm place, calculate the content (Pariza etc., 1999) of CLA in the fermented liquid according to typical curve.Reference: substratum adds and the linolic acid of sample with amount, does not connect bacterial classification, the same sample of other step.
1.5 strain identification
1.5.1 bacterium colony and morphologic observation: after the superior strain activation that obtains, be inoculated on the MR5 plate culture medium by four district's collimation methods, anaerobism is observed colony characteristics after cultivating 24h.Observe strain morphology feature, length range and width range etc. down in opticmicroscope behind the gramstaining.
1.5.2 Physiology and biochemistry is identified: with reference to " uncle's outstanding Bacteria Identification handbook the 9th edition (1994) is carried out experiments such as catalase reaction, gelatin liquification test, hydrogen sulfide reaction, indole reaction, temperature growth to the conjugated linolic acid superior strain that obtains and carried out preliminary evaluation.Produce evaluations of classifying of physiology such as acid, aerogenesis experiment and seminose, raffinose and biochemical aspect from glucose, experiment is with reference to " the 9th edition (1994) milk-acid bacteria classification evaluation of the outstanding Bacteria Identification handbook of uncle and experimental technique carry out concrete operations.
1.5.3 molecular biology identification: segmental recovery of purpose and purifying are all operated by the test kit description of step behind the pcr amplification of the preparation of template, bacterial strain 16SrDNA and the agarose gel electrophoresis.The purpose fragment that is recovered to is delivered to by precious biotechnology (Dalian) company limited then and checked order.With sequencing result use landed bacterium among BLAST software and the GenebanK the 16SrRNA sequence relatively, find and land of poor qualityly with the highest the landing the bacterial strain sequence number and access it of its similarity, carry out diversity ratio.
2. result
2.1 the screening of bacterial classification: when removing the cultivation of air conditions bottom fermentation, isolate 17 strains altogether and generate the stronger bacterial strain of CLA ability.By table 1 result as can be known, isolated bacterial strain CLA growing amount is between 4.316-33.442ug/mL (wet basis), and wherein No. 12 bacterial strain CLA output are up to 33.442ug/mL.This bacterial strain is named as ANCLA01.
2.2 strain identification:
2.2.1 the bacterium colony of bacterial classification and morphologic observation
Observe after the ANCLA01 bacterial strain MRS solid culture and find that bacterium colony projection, circle, smooth surface also are white in color; As shown in Figure 1, oily mirror is observed down and is found that the ANCLA01 bacterial strain is Gram-positive behind the gramstaining, short straight, the blunt circle in two ends of thalline, and no gemma, thalline is wide in 0.9～1.1um scope, and is long in 2～7um scope, becomes monomer, formation or short chain shape between thalline.Therefore, the ANCLA01 colonial morphology is with " the plant lactobacillus morphological specificity described in the outstanding Bacteria Identification handbook of uncle is more consistent.
2.2.2 Physiology and biochemistry check
Sugar fermentating test shows that ANCLA01 bacterial strain glucose fermentation result does not have gas and produces; Growth temperature detects finds that bacterial strain can be bred growth for 15 ℃, does not grow substantially for 45 ℃; Lactic acid generates and detects the test discovery, has lactic acid to generate in the fermented liquid.Take into account the above-mentioned bacterial strains morphological specificity, show this Pseudomonas homofermentation Type B lactobacillus.
2.2.2 molecular biology identification
The bacterial strain diversity ratio is found, 1482 base sequences identical (sequence has been uploaded GenBank, the number of landing EF185922) after 1482 base sequences that the 5th base of ANCLA01 bacterial strain 16SrRNA sequence is later and the 26th base of Lactobacillus plantarum strain L5.This result confirms that the ANCLA01 bacterial strain is strictly plant lactobacillus.
2.3 different grease additions cut up with a hay cutter ensiling raw materials such as corn stalk to 1～3cm to the influence of CLA output in the plant lactobacillus ANCLA01 ensiling, on one side the ensiling stockpile is pressed in silage tower, cellar for storing things or the bag, on one side by 8 * 10
7The bright sample of CFU/g sprays under the prerequisite of lactic acid nutrient solution (or the bright sample of 200g/t), the sunflower seed oil addition respectively by 2.0,3.0,4.0,5.0,6.0,7.0, the bright sample of 8.0kg/t (rapeseed oil or peanut oil addition by 5.0,6.0,7.0,8.0,9.0,10.0, the bright sample of 11.0kg/t) adds Vegetable oil lipoprotein, found that: when the bright sample of sunflower seed oil addition 5kg/t or rapeseed oil or peanut oil addition 8.0kg/t, CLA output is the highest, reaches 7.22mg/g (DW basis).The result shows that the sunflower seed oil optimum addition is the bright sample of 5kg/t, and rapeseed oil or peanut oil optimum addition are the bright sample of 8kg/t.
2.4. different lactobacillus inoculum amounts are to the influence of CLA output in the plant lactobacillus ANCLA01 ensiling
With ensiling raw materials such as corn stalk hand hay cutters to 1～3cm, the ensiling stockpile is pressed in silage tower, cellar for storing things or the bag, and meanwhile dosage press the bright sample of 5kg/t and add sunflower seed oil, one side inoculating lactic acid bacterium.Lactobacillus inoculum dosage is by 2 * 10
7CFU/g, 3 * 10
7CFU/g, 4 * 10
7CFU/g, 5 * 10
7CFU/g, 6 * 10
7CFU/g, 7 * 10
7The bright sample of CFU/g (or lactic acid bacteria culture solution 50,75,100,125,150, the bright sample of 175g/t), found that: the lactobacillus inoculum amount surpasses 4 * 10
7During CFU/g aquatic foods sample, CLA output no longer obviously increases after reaching 8.84mg/g (DW basis) in the ensiling, shows that the milk-acid bacteria optimum inoculation amount is 4-6 * 10
7The bright sample of CFU/g (or the bright sample of lactic acid bacteria culture solution 100-150g/t).
Utilize this plant lactobacillus fermentative preparation to be rich in the method for conjugated linolic acid silage:
1) plant lactobacillus ANCLA01 bacterial strain being carried out batch spreads cultivation: after will preserving twice of bacterial strain activation with above-mentioned MRS liquid nutrient medium, be inoculated in the 250mL triangular flask, every bottled liquid measure 100mL is after 37 ℃ of constant temperature leave standstill and cultivate 48～72h, again by 2 * 10
7CFU/mL adds concentration and adds in the liquid seed fermentation jar, mixes back cultivation and fermentation 48h under 37 ℃ of conditions.
2) inoculation plant lactobacillus ANCLA01 in silage: ensiling raw materials such as corn stalk are cut up with a hay cutter to 1～3cm, on one side the ensiling stockpile is pressed in silage tower, cellar for storing things or the bag, spray plant lactobacillus ANCLA01 liquid and Vegetable oil lipoprotein after above-mentioned spreading cultivation on one side; Plant lactobacillus ANCLA01 dosage of inoculation is pressed 4-6 * 10
7The bright sample of CFU/g (or the lactic acid bacteria culture solution addition is the bright sample of 100-150g/t), sunflower seed oil addition are the bright sample of 5kg/t (rapeseed oil or peanut oil addition are the bright sample of 8kg/t); At last with silage tower, cellar for storing things or bag compacting sealing; Can enable behind the ensiling spontaneous fermentation 40-60d, more than actual measurement CLA content is all above 7.52mg/g (DW basis).
Claims (6)
1, a kind of plant lactobacillus ANCLA01 is characterized in that described bacterial strain is Lactobacillusplantarum ANCLA01, and this bacterial strain preservation is registered on the books is numbered CGMCCNo.1906.
2, a kind of plant lactobacillus fermentative preparation is rich in the method for conjugated linolic acid silage, it is characterized in that:
1) plant lactobacillus ANCLA01 bacterial strain being carried out batch spreads cultivation: after will preserving twice of bacterial strain activation with the MRS liquid nutrient medium, be inoculated in the 250mL triangular flask, and every bottled liquid measure 100mL, 37 ℃ of constant temperature press 2 * 10 after leaving standstill and cultivating 48～72h again
7CFU/mL adds concentration and adds in the liquid seed fermentation jar, mixes back cultivation and fermentation 48h under 37 ℃ of conditions;
2) inoculation plant lactobacillus ANCLA01 in silage: on one side the ensiling stockpile is pressed in silage tower, cellar for storing things or the bag, on one side plant lactobacillus ANCLA01 liquid and the Vegetable oil lipoprotein of sprinkling after above-mentioned spreading cultivation; At last with silage tower, cellar for storing things or bag compacting sealing; Can enable behind the ensiling spontaneous fermentation 40-60d.
3, a kind of plant lactobacillus fermentative preparation according to claim 2 is rich in the method for conjugated linolic acid silage, it is characterized in that: described plant lactobacillus ANCLA01 dosage of inoculation is pressed 4-6 * 10
7Bright sample of CFU/g or lactic acid bacteria culture solution addition are the bright sample of 100-150g/t.
4, a kind of plant lactobacillus fermentative preparation according to claim 2 is rich in the method for conjugated linolic acid silage, it is characterized in that: described Vegetable oil lipoprotein is sunflower seed oil, rapeseed oil or peanut oil.
5, a kind of plant lactobacillus fermentative preparation according to claim 4 is rich in the method for conjugated linolic acid silage, it is characterized in that: described sunflower seed oil addition is the bright sample of 5kg/t.
6, a kind of plant lactobacillus fermentative preparation according to claim 4 is rich in the method for conjugated linolic acid silage, it is characterized in that: described rapeseed oil or peanut oil addition are the bright sample of 8kg/t.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2007100196553A CN101020895B (en) | 2007-01-24 | 2007-01-24 | Method for preparing silage rich in conjugated linoleic acid by lactobacillus plantarum fermentation |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2007100196553A CN101020895B (en) | 2007-01-24 | 2007-01-24 | Method for preparing silage rich in conjugated linoleic acid by lactobacillus plantarum fermentation |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN101020895A true CN101020895A (en) | 2007-08-22 |
| CN101020895B CN101020895B (en) | 2011-05-04 |
Family
ID=38708771
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN2007100196553A Expired - Fee Related CN101020895B (en) | 2007-01-24 | 2007-01-24 | Method for preparing silage rich in conjugated linoleic acid by lactobacillus plantarum fermentation |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN101020895B (en) |
Cited By (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101449752B (en) * | 2007-11-30 | 2011-04-27 | 河南农业大学 | Composite microbial inoculant for corn silage and using method thereof |
| CN101074426B (en) * | 2007-01-24 | 2011-06-29 | 合肥市科茂隆生物工程有限公司 | Method for producing bean-dregs feed containing conjugated linolic acid by plant lactobacillin fermentation |
| CN102250807A (en) * | 2011-07-06 | 2011-11-23 | 广西工学院 | Microbial agent for Chinese silvergrass ensilage as well as preparation method and application thereof |
| CN101632411B (en) * | 2009-08-20 | 2012-07-11 | 陈志敏 | Method for preparing high-quality rapeseed protein containing conjugated linoleic acid by multi-strain fermentation |
| CN103719551A (en) * | 2013-12-27 | 2014-04-16 | 柳州市青钱柳农业科技开发有限公司 | Preparation method of GABA (gamma-aminobutyric acid)-rich silage |
| CN104336416A (en) * | 2014-11-03 | 2015-02-11 | 郑州大学 | Lactobacillus plantarum and application thereof to alfalfa silage |
| CN107232409A (en) * | 2017-08-01 | 2017-10-10 | 合肥合丰牧业有限公司 | A kind of high-quality meat sheep feed |
| CN115960767A (en) * | 2022-11-09 | 2023-04-14 | 厦门元之道生物科技有限公司 | Lactobacillus plantarum and application thereof |
| CN118685316A (en) * | 2024-07-03 | 2024-09-24 | 南昌大学 | A strain of Lactobacillus plantarum R6 and its application |
Family Cites Families (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN100342005C (en) * | 2005-11-07 | 2007-10-10 | 浙江工商大学 | Process for preparing high actived alctic acid bacteria product |
| CN100396781C (en) * | 2005-11-07 | 2008-06-25 | 浙江工商大学 | Method for preparing conjugated linoleic acid by using lactic acid bacteria |
| CN101074426B (en) * | 2007-01-24 | 2011-06-29 | 合肥市科茂隆生物工程有限公司 | Method for producing bean-dregs feed containing conjugated linolic acid by plant lactobacillin fermentation |
-
2007
- 2007-01-24 CN CN2007100196553A patent/CN101020895B/en not_active Expired - Fee Related
Cited By (12)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101074426B (en) * | 2007-01-24 | 2011-06-29 | 合肥市科茂隆生物工程有限公司 | Method for producing bean-dregs feed containing conjugated linolic acid by plant lactobacillin fermentation |
| CN101449752B (en) * | 2007-11-30 | 2011-04-27 | 河南农业大学 | Composite microbial inoculant for corn silage and using method thereof |
| CN101632411B (en) * | 2009-08-20 | 2012-07-11 | 陈志敏 | Method for preparing high-quality rapeseed protein containing conjugated linoleic acid by multi-strain fermentation |
| CN102250807A (en) * | 2011-07-06 | 2011-11-23 | 广西工学院 | Microbial agent for Chinese silvergrass ensilage as well as preparation method and application thereof |
| CN102250807B (en) * | 2011-07-06 | 2013-06-19 | 广西工学院 | Microbial agent for Chinese silvergrass ensilage as well as preparation method and application thereof |
| CN103719551A (en) * | 2013-12-27 | 2014-04-16 | 柳州市青钱柳农业科技开发有限公司 | Preparation method of GABA (gamma-aminobutyric acid)-rich silage |
| CN103719551B (en) * | 2013-12-27 | 2015-09-16 | 柳州市青钱柳农业科技开发有限公司 | A kind of preparation method being rich in the ensilage of GABA |
| CN104336416A (en) * | 2014-11-03 | 2015-02-11 | 郑州大学 | Lactobacillus plantarum and application thereof to alfalfa silage |
| CN107232409A (en) * | 2017-08-01 | 2017-10-10 | 合肥合丰牧业有限公司 | A kind of high-quality meat sheep feed |
| CN115960767A (en) * | 2022-11-09 | 2023-04-14 | 厦门元之道生物科技有限公司 | Lactobacillus plantarum and application thereof |
| CN115960767B (en) * | 2022-11-09 | 2024-04-26 | 厦门元之道生物科技有限公司 | Lactobacillus plantarum and application thereof |
| CN118685316A (en) * | 2024-07-03 | 2024-09-24 | 南昌大学 | A strain of Lactobacillus plantarum R6 and its application |
Also Published As
| Publication number | Publication date |
|---|---|
| CN101020895B (en) | 2011-05-04 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN101074426B (en) | Method for producing bean-dregs feed containing conjugated linolic acid by plant lactobacillin fermentation | |
| CN101020895A (en) | Method for preparing silage rich in conjugated linoleic acid by lactobacillus plantarum fermentation | |
| CN103627662B (en) | A kind of Bradyrhizobium sp Arachis and uses thereof | |
| CN103468594B (en) | Candidautilis strain and application thereof | |
| CN107227277A (en) | A kind of Lactobacillus plantarum E680 and its application | |
| CN110063406A (en) | Bacillus amyloliquefaciens and its fermentation seed liquid, application and soybean meal fermenting method | |
| CN100569704C (en) | A K3 strain capable of dissolving calcium phosphate in soil and its microbial organic fertilizer | |
| CN101696395A (en) | Antagonistic bacterium for preventing and killing off continuous cropping tobacco black shank and microbial organic fertilizer thereof | |
| CN100469890C (en) | Preparation method of conjugated linoleic acid | |
| CN102559552A (en) | Production method and application of high-yield gamma-aminobutyric acid | |
| CN113980853B (en) | Lactic acid-producing lactococcus garvieae WBT0008 and application thereof | |
| CN107354102B (en) | A high-sugar-tolerant Pichia pastoris strain and its application | |
| CN105132321A (en) | Enterococcus faecium, culture medium thereof for high-density solid-state fermentation, and high-density solid-state fermentation method | |
| CN101632411B (en) | Method for preparing high-quality rapeseed protein containing conjugated linoleic acid by multi-strain fermentation | |
| CN103980047B (en) | Microbial manure of a kind of water resistant rice sprouts damping-off and preparation method thereof | |
| CN106566788B (en) | Utilize the method and its application of resistance to silver-colored Soil Bacillus biosynthesis nanometer selenium | |
| CN101914477A (en) | A strain of Lactobacillus plantarum and its use | |
| CN115820464A (en) | Paenibacillus cellularis JAUCCB0001 and application thereof in promoting growth of oudemansiella radicata | |
| CN111154692B (en) | A strain of Lactobacillus plantarum that degrades nitrite and inhibits ACE | |
| CN110713956B (en) | Lysine bacillus S12 and application thereof | |
| CN117448209B (en) | Lactobacillus pentosus for efficiently degrading phosphorus phytate, fermentation preparation and application thereof | |
| CN102078027B (en) | Method for separating microorganisms on surface of tobacco leaf at high temperature by utilizing pure tobacco leaf leach liquor solid plate | |
| CN110452849B (en) | A kind of probiotic Lactobacillus plantarum | |
| CN107118885A (en) | A kind of method that the fermented wine containing GABA is produced using the piece of resistance to ethanol coccus | |
| CN107034161A (en) | It is a kind of to produce Lactobacillus kefiranofaciens and its application in capsicum is fermented |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant | ||
| EE01 | Entry into force of recordation of patent licensing contract |
Assignee: The very fast epoch of the Huaibei Animal nutrition Co., Ltd Assignor: Anhui Agricultural University Contract record no.: 2012340000016 Denomination of invention: Plant lactobacillus fermenting process for preparing ensilage with rich conjugated linoleic acid Granted publication date: 20110504 License type: Exclusive License Open date: 20070822 Record date: 20120315 |
|
| C17 | Cessation of patent right | ||
| CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20110504 Termination date: 20140124 |